Analysis of glycans derived from glycoconjugates by capillary electrophoresis-mass spectrometry.
Analysis of glycans derived from glycoconjugates by capillary electrophoresis-mass spectrometry.
复制标题
通过毛细管电泳质谱法衍生出糖缀合物的聚糖分析。
DOI：
10.1002/elps.201100342
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发表时间：
2011-12
期刊：
影响因子：
2.9
通讯作者：
Mechref Y
中科院分区：
文献类型：
--
作者：
Mechref Y
衍生自糖缀合物的聚糖的高度结构变化（其随着蛋白质的分子大小而显著增加）有助于通常与糖缀合物相关的糖基化模式的复杂性。在糖蛋白的情况下，这种变异源于蛋白质的多个糖基化位点和与每个位点相关的聚糖结构的数量（微异质性）。全面表征高度复杂的聚糖混合物的能力在分析上具有刺激性和挑战性。尽管最强大的质谱（MS）和串联MS技术能够提供丰富的结构信息，但在没有高阶串联MS（MSn）的情况下，它们仍然无法轻易识别异构聚糖结构。已使用几种分离方法实现异构聚糖结构的分析，包括高pH阴离子交换色谱法（HPAEC）、亲水相互作用色谱法（HILIC）和气相色谱法（GC）。然而，毛细管电泳（CE）和微流体毛细管电泳（MCE）提供高分离效率和分辨率，允许分离密切相关的聚糖结构。因此，将CE和MCE连接到MS是一种强大的分析方法，可以对复杂聚糖样品进行潜在的全面和灵敏的分析。本文综述了不同的CE和MCE方法在糖蛋白结构表征中的实用性，以及将这些方法与质谱连接的可行性。
The high structural variation of glycan derived from glycoconjugates, which substantially increases with the molecular size of a protein, contributes to the complexity of glycosylation patterns commonly associated with glycoconjugates. In the case of glycoproteins, such variation originates from the multiple glycosylation sites of proteins and the number of glycan structures associated with each site (microheterogeneity). The ability to comprehensively characterize highly complex mixture of glycans has been analytically stimulating and challenging. Although the most powerful mass spectrometric (MS) and tandem MS techniques are capable of providing a wealth of structural information, they are still not able to readily identify isomeric glycan structures without high order tandem MS (MSn). The analysis of isomeric glycan structures has been attained using several separation methods, including high-pH anion exchange chromatography (HPAEC), hydrophilic interaction chromatography (HILIC) and gas chromatography (GC). However, capillary electrophoresis (CE) and microfluidics capillary electrophoresis (MCE) offer high separation efficiency and resolutions, allowing the separation of closely related glycan structures. Therefore, interfacing CE and MCE to MS is a powerful analytical approach, allowing potentially comprehensive and sensitive analysis of complex glycan samples. This review describes and discusses the utility of different CE and MCE approaches in the structural characterization of glycoproteins and the feasibility of interfacing these approaches to mass spectrometry.
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